LOCUS       sequence-181902-        3105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module B, Promoter: TaU6, Gene: Esp3I ccdb cassette for single gRNA 
            spacer cloning, Terminator: Pol III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3105)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 3105)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pMOD_B2518
FEATURES             Location/Qualifiers
     source          1..3105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      80..668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     569..588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     822..839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1240..1545)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(1342..1361)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1872..1947
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2013..2117
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
       61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
      121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt
      181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc
      421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      841 aagcggaaga gcgcccacct gccaggggac gaccaagccc gttattctga cagttctggt
      901 gctcaacaca tttatattta tcaaggagca cattgttact cactgctagg agggaatcga
      961 actaggaata ttgatcagag gaactacgag agagctgaag ataactgccc tctagctctc
     1021 actgatctgg gtcgcatagt gagatgcagc ccacgtgagt tcagcaacgg tctagcgctg
     1081 ggcttttagg cccgcatgat cgggcttttg tcgggtggtc gacgtgttca cgattgggga
     1141 gagcaacgca gcagttcctc ttagtttagt cccacctcgc ctgtccagca gagttctgac
     1201 cggtttataa actcgcttgc tgcatcagac ttggagacgt tatattcccc agaacatcag
     1261 gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat
     1321 aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat
     1381 atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg cactggccag
     1441 ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag
     1501 acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt
     1561 tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct
     1621 gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc
     1681 ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc
     1741 aactgtcact gtaatacgct gcttcatagc atacctcctt ttgacatact tcgggtatac
     1801 atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg
     1861 gcttcgtctc agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     1921 cttgaaaaag tggcaccgag tcggtgcttt ttttccggtg acgcaggtga tgaaaagcct
     1981 tagggtcagg tggcactttt cggggaaatg tgcgatgaac ccctatttgt ttatttttct
     2041 aaatacattc aaatatgtat ccgctaatga gacaataacc ctgataaatg cttcaattat
     2101 attgaaaaag gaagagtatg agtatccaac atttccgtgt cgcccttatt cccttttttg
     2161 cggccttctg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2221 aagatcagtt gggtgcccga gtgggttaca ttgaactgga tctcaacagc ggcaaaatct
     2281 tagagagttt tcgccccgaa gaacgcttcc caatgatgag cactttcaaa gttctgctat
     2341 gtggcgcggt gttatcccgt attgatgctg ggcaagagca actcggtcgc cgcatacact
     2401 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     2461 tgacagtaag agaattatgt agtgctgcca taaccatgag tgataacact gcggccaact
     2521 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2641 agcgtgacac cacgatgcct gtagcaatgg caacaactct ccgcaaacta ttaactggcg
     2701 aactacttac tcttgcttca cgccagcaac tcattgactg gatggaggcg gacaaagttg
     2761 caggaccact tctgcgctcg gcacttccgg ctggctggtt tattgctgat aaatctggag
     2821 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2941 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagacgaa gggaaataaa
     3001 tagtagcccg cctgatgtgc gggctttttt ttggtggtgg ttaccagtgt cgcttagcta
     3061 ccgtaggtcc tcttaggcgg tccgattagc gacatagtca tcaat
//
